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Acylated Uridine and Cytidine and Uses Thereof

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Acylated Uridine and Cytidine and Uses Thereof ~" ' Nil II II II INI II MM INI Ml Ml J European Patent Office _ __ © Publication number: 0 339 075 B1 Office_„. europeen des brevets © EUROPEAN PATENT SPECIFICATION © Date of publication of patent specification: 18.08.93 © Int. CI.5: C07H 19/067, A61 K 31/70 © Application number: 88909932.1 @ Date of filing: 27.10.88 © International application number: PCT/US88/03823 © International publication number: WO 89/03837 (05.05.89 89/10) &) ACYLATED URIDINE AND CYTIDINE AND USES THEREOF. ® Priority: 28.10.87 US 115929 © Proprietor: PRO-NEURON, INC. 1530 East Jefferson Street @ Date of publication of application: Rockvllle, MD 20852(US) 02.11.89 Bulletin 89/44 @ Inventor: VON BORSTEL, Reld, Warren © Publication of the grant of the patent: 3115 University Blvd. West 18.08.93 Bulletin 93/33 Kensington, MD 20895(US) Inventor: BAMAT, Michael, Kevin © Designated Contracting States: 6516 Western Avenue AT BE CH DE FR GB IT LI LU NL SE Chevy Chase, MD 2081 5(US) References cited: EP-A- 0 056 265 EP-A- 0 178 267 © Representative: Rltter, Stephen David et al EP-A- 0 222 192 JP-A- 5 223 085 Mathys & Squire 10 Fleet Street JP-A- 5 524 150 JP-A- 5 791 995 London EC4Y 1AY (GB) JP-A- 5 849 315 US-A- 3 585 188 US-A- 3 894 000 US-A- 3 975 367 US-A- 3 991 045 CHEMICAL ABSTRACTS, vol. 74, no. 21, 24 00 May 1971, Columbus.OH (US); RAJABELEE, p. 433, no. 112368k# oo Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid (Art. 99(1) European patent convention). Rank Xerox (UK) Business Services (3. 10/3.6/3.3. 1) EP 0 339 075 B1 Description This application is a continuation-in-part of United States application Ser. No. 115,929, filed October 28, 1987, the disclosure of which is incorporated herein by reference. 5 Field of The Invention This invention relates generally to acyl derivatives of cytidine and uridine and to the use of those derivatives to deliver exogenous ribonucleosides to animal tissue. More specifically, this invention relates to io the acyl derivatives of cytidine and uridine, and the uses of those novel derivatives to deliver these ribonucleosides to animal tissue and thereby to support cellular metabolic functions. The novel acyl derivatives and mixtures thereof may be used to treat or prevent a variety of physiological and pathological conditions, including treatment of liver disease or damage, cerebrovascular disorders, respiratory distress syndromes, cardiac damage, and other clinical conditions. 75 Background of The Invention There are many physiological and pathological conditions of animal tissue where the supply of exogenous ribonucleosides nay have useful therapeutic applications. In a number of physiological and 20 pathological conditions, the administration to an animal of RNA, nucleotides, or individual or mixtures of nucleosides, has been shown to improve the natural repair processes of the affected cells. There are many important metabolic reactions that are usually functionally subsaturated and limited by availability of either substrates or cofactors. Such rate-limiting compounds may be either nutritionally essential or synthesized de novo in the body. Under conditions of tissue trauma, infection or adaptation to 25 physiological demand, particularly when cellular repair or regeneration processes are activated, the optimum nutritional, biochemical, or hormonal environment for promoting such repair may be quite different from the requirements for normal cell and tissue function. In such cases, therapeutic benefit may be derived by providing appropriate conditionally essential nutrients, such as ribonucleosides or metabolites which may be required in quantities not usually available from a normal diet. The therapeutic potential for this strategy 30 of directly supporting the metabolic function of damaged or diseased tissues has not been realized in contemporary medical practice. At the cellular level of organization, there are specific metabolic responses to trauma that are involved, in a variety of tissues, in the processes of tissue repair, regeneration, or adaptation to altered functional demand. Most processes of tissue damage and repair are accompanied by a substantial increase in the 35 activity of the hexose monophosphate pathway of glucose metabolism. The hexose monophosphate pathway is the route of formation for the pentose sugars (e.g., ribose) which are necessary for nucleotide and nucleic acid synthesis. The availability of ribose is rate limiting for nucleotide synthesis under most physiological or pathological conditions. Rapid production of nucleotides for the synthesis of nucleic acids and nucleotide-derived cofactors (such as cytidine di-phosphocholine 40 (CDP choline) or uridine di-phosphoglucose (UDPG)) is essential for the processes of tissue repair and cellular proliferation. Even though nucleotides are synthesized de novo from simpler nutrients, so that there is not an absolute dietary requirement for direct nucleotide precursors, many tissues may not have optimal capacity for nucleotide synthesis particularly during tissue repair or cellular proliferation. It is possible to bypass the limited capacity of the hexose monophosphate pathway by providing 45 preformed ribonucleosides directly to tissues where they are incorporated in the nucleotide pools via the "salvage" pathways of nucleotide synthesis. It is also possible that pyrimidine ribonucleosides may exert therapeutic influences through mechanisms unrelated to the support of nucleotide biosynthesis. The effects of the administration of pyrimidine nucleosides, and in particular, uridine and cytidine, on a variety of physiological and pathological conditions in experimental animals, isolated tissues, and to some 50 extent, in humans, have been extensively studied. These are summarized below. (1) Heart In isolated rat hearts subjected to low-flow ischemia, reperfusion with uridine induced restoration of 55 myocardial ATP levels, total adenine nucleotide content, uridine nucleotide levels, and glycogen content. Ischemia was reported to produce a breakdown of creatinine phosphate, ATP, uridine nucleotides and glycogen. Aussedat, J., Cardiovasc. Res. 17:145-151 (1983). 2 EP 0 339 075 B1 In a related study, perfusion of isolated rat hearts with uridine resulted in a concentration-dependent elevation of myocardial uracil nucleotide content. Following low-flow ischemia, the rate of incorporation of uridine was increased two fold. Aussedat, J., et al., Mol. Physiol. 6:247-256 (1984). In another study, isoproterenol was administered to rats which depleted cardiac glycogen stores and 5 reduced myocardial UTP and UDP-glucose levels. Despite the spontaneous restoration of myocardial UTP levels, UDP-glucose concentrations remained depressed unless uridine or ribose were administered. Prolonged intravenous infusion of ribose or uridine resulted in a restoration of myocardial glycogen. Thus, there may be compartmentation of uridine nucleotides in the heart, with the pools being fed differentially by the salvage or de novo pathways of pyrimidine synthesis. Aussedat, J., et al., J. Physiol. 78:331-336 (1982). io The effects of nucleosides on acute left ventricular failure in isolated dog heart was studied by Buckley, N.M., et al., Circ. Res 7:847-867 (1959). Left ventricular failure was induced in isolated dog hearts by increasing aorta pressure. In this model, guanosine, inosine, uridine and thymidine were found to be positive inotropic agents, while cytidine and adenosine were negatively inotropic. Sodium uridine monophosphate (UMP) and potassium orotate were found to increase the animal's is resistance to subsequent adrenaline-induced myocardial necrosis. These compounds reduced mortality and improved myocardial function as assessed by ECG readings, biochemical findings, and relative heart weight. Intravenous administration of UMP exerted a more pronounced prophylactic effect than did potassium orotate. Kuznetsova, L.V., et al., Farmakol.-Toksikol 2: 170-173 (1981). In a study on the effects of hypoxia in isolated rabbit hearts, myocardial performance declined while 20 glucose uptake with glycolysis, glycogenolysis and breakdown of adenine nucleotides were reportedly increased. Administration of uridine increased myocardial performance, glucose uptake and glycolysis and also diminished the disappearance of glycogen and adenine nucleotides from hypoxic hearts. Uridine also increased glucose uptake, glycolysis, levels of ATP and glycogen, as well as myocardial performance in propranolol-treated hearts. Kypson, J., et al., J. Mol. Cell. Cardiol. 10:545-565 (1978). 25 In a study of pyrimidine nucleotide synthesis from exogenous cytidine in the isolated rat heart, myocardial cytosine nucleotide levels were significantly increased by a 30 minute supply of cytidine. Most of the cytidine was recovered as part of cytosine nucleotides and uracil nucleotides. Very little of the cytidine that was taken up was converted into uridine nucleotides. These results suggest that the uptake of cytidine can play an important part in myocardial cytosine nucleotide metabolism. Lortet, S., et al., Basic 30 Res. Cardiol. 81:303-310 (1986). In another study, myocardial fatigue was produced by repeated, brief ligations of the ascending aorta. Intravenous administration of a mixture of
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